Η συμμετοχή των Τ-Ρυθμιστικών Λεμφοκυττάρων στην παθογένεια της σηπτικού συνδρόμου

Εισαγωγή: Οι υποπληθυσμοί των μονοκυττάρων λειτουργούν ως πρώτη γραμμή άμυνας του ξενιστή. Η σχέση των απόλυτων αριθμών τους με την τελική έκβαση των ανοσοκατεσταλμένων ξενιστών δεν έχει κατανοηθεί πλήρως μέχρι στιγμής. Μέθοδοι: Στο πρώτο στάδιο της μελέτης μετρήθηκαν με κυτταρομετρία ροής οι απόλυτοι αριθμοί των κυκλοφορούντων CD14θετ/HLA-DRθετ/CD45θετ μονοκυττάρων σε 70 ασθενείς με λοιμώξεις από Gram-αρνητικούς μικροοργανισμούς και σε 10 υγιείς εθελοντές. Στη δεύτερη φάση της μελέτης προσδιορίσθηκαν ανοσοφαινοτυπικά οι απόλυτοι αριθμοί των CD14θετ/CD16αρν/CD45θετ και των CD14θετ/CD16θετ/CD45θετ κυκλοφορούντων φλεγμονωδών μονοκυττάρων καθώς και των CD14ασθ/CD16θετ/CD45θετ περιπολούντων μονοκυττάρων σε άλλους 55 ασθενείς και 10 υγιείς εθελοντές. Οι μετρήσεις επαναλήφθηκαν τις ημέρες 3, 7 και 10. Τα αποτελέσματα συσχετίσθηκαν με την επιβίωση μετά από 28 ημέρες. Αποτελέσματα: Υψηλότεροι αριθμοί HLA-DR θετικών μονοκυττάρων την ημέρα 1 και την ημέρα 3 ανευρέθηκαν σε αυτούς που επιβίωσαν σε σχέση με αυτούς που δεν επιβίωσαν (p=0.030) και τους υγιείς εθελοντές (p<0.0001). Σε όσους δεν επιβίωσαν οι απόλυτοι αριθμοί δε διέφεραν από τους υγιείς εθελοντές. Η ανάλυση ROC κατέδειξε ότι όριο υψηλότερο από 373 κύτταρα/mm3 την ημέρα 1 θα μπορούσε να διακρίνει αυτούς που επιβίωσαν από αυτούς που δεν επιβίωσαν. Αυτό σχετιζόταν με σχετικό κίνδυνο (OddsRatio, OR) 4.82 για δυσμενή έκβαση (p=0.020). Οι απόλυτοι αριθμοί των περιπολούντων μονοκυττάρων ήταν υψηλότεροι σε όσους επιβίωσαν σε σύγκριση με όσους δεν επιβίωσαν τις ημέρες 3 και 7. Η ανάλυση ROCέδειξε ότι το όριο των 27 κυττάρων/mm3 την ημέρα 3 θα μπορούσε να διακρίνει αυτούς που επιβίωσαν από αυτούς που δεν επιβίωσαν, προστατεύοντας από τον θάνατο (OR0.09, p=0.031). Από την ανάλυση λογιστικής παλινδρόμησης φάνηκε ότι η παρουσία πυελονεφρίτιδας αποτελεί ανεξάρτητη μεταβλητή που σχετίζεται με την τελική έκβαση (OR 2.29, p=0.002).Background: Evidence on the changes in the absolute counts of monocyte subpopulations in sepsis is missing. Methods: Firstly, absolute counts of circulating CD14pos/HLA-DRpos/CD45pos monocytes were measured by flow cytometry in 70 patients with Gram-negative sepsis and in 10 healthy volunteers. In the second phase, immunophenotyping was performed and the absolute count of circulating inflammatory monocytes and of circulating CD14dim/CD16pos/CD45pos patrolling monocytes were measured in another 55 patients and 10 healthy volunteers. Measurements were repeated on days 3, 7, and 10. Results were correlated with survival after 28 days. Results: Greater numbers of CD14pos/HLA-DRpos/CD45pos monocytes were found on day 1 in survivors compared to nonsurvivors (p = 0.030). Receiver operating characteristic (ROC) analysis showed that a cutoff higher than 337 cells/mm3 on day 1 could discriminate between survivors and nonsurvivors with a positive predictive value (PPV) of 91.1%. Logistic regression including Sequential Organ Failure Assessment (SOFA) score and Acute Physiology and Chronic Health Evaluation (APACHE) II score showed that an absolute count greater than 337 cells/mm3 was independently associated with unfavorable outcome (odds ratio (OR) 0.19, p = 0.050). The absolute counts of inflammatory and of CD14dim/CD16pos/CD45pos monocytes were greater in patients than healthy controls during the entire 10 days of follow-up. The absolute counts on day 3 of CD14dim/CD16pos/CD45pos monocytes were greater in survivors than nonsurvivors (p = 0.027). ROC analysis revealed that the cutoff at 27 cells/mm3 could discriminate between survivors and nonsurvivors with PPV of 94.1%. Logistic regression including age, SOFA score, and APACHE II score showed that an absolute count greater than 27 cells/mm3 was independently associated with unfavorable outcome (OR 0.06, p = 0.033). Logistic regression analysis showed that intra-abdominal infection on day 1 was predictive of low CD14dim/ CD16pos/CD45pos count on day 3. Conclusion: Circulating counts of inflammatory and patrolling monocytes are greatly increased in Gram-negative sepsis. Absolute counts of CD14pos/HLA-DRpos/CD45pos monocytes on day 1 and CD14dim/CD16pos/CD45pos monocytes on day 3 are independently associated with final outcome

Increases in inflammatory and CD14dim/CD16pos/CD45pos patrolling monocytes in sepsis: correlation with final outcome

Abstract Background Evidence on the changes in the absolute counts of monocyte subpopulations in sepsis is missing. Methods Firstly, absolute counts of circulating CD14pos/HLA-DRpos/CD45pos monocytes were measured by flow cytometry in 70 patients with Gram-negative sepsis and in 10 healthy volunteers. In the second phase, immunophenotyping was performed and the absolute count of circulating inflammatory monocytes and of circulating CD14dim/CD16pos/CD45pos patrolling monocytes were measured in another 55 patients and 10 healthy volunteers. Measurements were repeated on days 3, 7, and 10. Results were correlated with survival after 28 days. Results Greater numbers of CD14pos/HLA-DRpos/CD45pos monocytes were found on day 1 in survivors compared to nonsurvivors (p = 0.030). Receiver operating characteristic (ROC) analysis showed that a cutoff higher than 337 cells/mm3 on day 1 could discriminate between survivors and nonsurvivors with a positive predictive value (PPV) of 91.1%. Logistic regression including Sequential Organ Failure Assessment (SOFA) score and Acute Physiology and Chronic Health Evaluation (APACHE) II score showed that an absolute count greater than 337 cells/mm3 was independently associated with unfavorable outcome (odds ratio (OR) 0.19, p = 0.050). The absolute counts of inflammatory and of CD14dim/CD16pos/CD45pos monocytes were greater in patients than healthy controls during the entire 10 days of follow-up. The absolute counts on day 3 of CD14dim/CD16pos/CD45pos monocytes were greater in survivors than nonsurvivors (p = 0.027). ROC analysis revealed that the cutoff at 27 cells/mm3 could discriminate between survivors and nonsurvivors with PPV of 94.1%. Logistic regression including age, SOFA score, and APACHE II score showed that an absolute count greater than 27 cells/mm3 was independently associated with unfavorable outcome (OR 0.06, p = 0.033). Logistic regression analysis showed that intra-abdominal infection on day 1 was predictive of low CD14dim/ CD16pos/CD45pos count on day 3. Conclusion Circulating counts of inflammatory and patrolling monocytes are greatly increased in Gram-negative sepsis. Absolute counts of CD14pos/HLA-DRpos/CD45pos monocytes on day 1 and CD14dim/CD16pos/CD45pos monocytes on day 3 are independently associated with final outcome. Trial registration ClinicalTrials.gov, NCT01223690. Registered retrospectively on 18 October 2010

Additional file 2: of Increases in inflammatory and CD14dim/CD16pos/CD45pos patrolling monocytes in sepsis: correlation with final outcome

Figure S1. Absolute counts of circulating CD14pos/HLA-DRpos/CD45pos monocytes and subpopulations of monocytes in relation to treatment allocation. Absolute counts of (A) CD14pos/HLA-DRpos/CD45pos monocytes, (B) inflammatory monocytes, and (C) CD14dim/CD16pos/CD45pos patrolling monocytes on days 1 and 3 between patients allocated to treatment with placebo and patients allocated to treatment with clarithromycin. P values refer to the indicated comparisons. (DOCX 105 kb

Additional file 1: of Increases in inflammatory and CD14dim/CD16pos/CD45pos patrolling monocytes in sepsis: correlation with final outcome

Table S1. Demographic characteristics of patients in both study phases. (DOCX 16 kb

Additional file 4: of Increases in inflammatory and CD14dim/CD16pos/CD45pos patrolling monocytes in sepsis: correlation with final outcome

Table S3. Differences of baseline characteristics of day 1 between survivors and nonsurvivors of the second phase of the study. (DOCX 18 kb